I stained cells with different monoclonal antibodies. Unfortunately after staining the cells, our flow cytometer had broken down. I had the cells pulled and cryopreserved. I wonder would I be able to stain the cells with different monoclonal antibodies again after thawing them later?
Thx

So, you stained cells for flow, and freeze the cell?
my questin is your stained cell still alive? or you fixed?
if cell is still viable you can stain again with different antibody, no problem.
if you fixed the cell, prob not.

I stained cells with different monoclonal antibodies. Unfortunately after staining the cells, our flow cytometer had broken down. I had the cells pulled and cryopreserved. I wonder would I be able to stain the cells with different monoclonal antibodies again after thawing them later?
Thx

Yes. I stained the cell for flow cytometry analysis, I freeze them up becoz our flow cytometer broken down.
The cells are still alive, I didn't fix them.
I need to stain them again with the same set of monoclonal antibodies. Just wondering whether these cells can still pick up the monoclonal antibodiyes as their receptors might be taken up by the previous staining.
Thx.

So, you stained cells for flow, and freeze the cell?
my questin is your stained cell still alive? or you fixed?
if cell is still viable you can stain again with different antibody, no problem.
if you fixed the cell, prob not.

QUOTE (yftan2 @ Mar 5 2008, 05:32 PM)

I stained cells with different monoclonal antibodies. Unfortunately after staining the cells, our flow cytometer had broken down. I had the cells pulled and cryopreserved. I wonder would I be able to stain the cells with different monoclonal antibodies again after thawing them later?
Thx

well, I never done what you did for your cells, so this is just my assumption

if the cell is not fixed and alive, after certain time you staied with antibody, cell will do so called "capping"
capping is basically cell realize that some antibody attached to the protein so cell will internalize the protein with antibody and digest them.

so if capping happen before you freeze, I think your cell is back to original condition (before you stain with antibody).

However, I am not sure the effect of freezing to the expression of the antigen to your antibody,
on the other hands, Freezing decrease expression of certain protein that you want to detect by your antibody, you might get unexpected result.

again, those are just my opinion, since I never freeze stained cell, so I hope it help, but if this is not what you are looking for, than I am sorry

Yes. I stained the cell for flow cytometry analysis, I freeze them up becoz our flow cytometer broken down.
The cells are still alive, I didn't fix them.
I need to stain them again with the same set of monoclonal antibodies. Just wondering whether these cells can still pick up the monoclonal antibodiyes as their receptors might be taken up by the previous staining.
Thx.

QUOTE (Rnotk @ Mar 7 2008, 02:05 AM)

So, you stained cells for flow, and freeze the cell?
my questin is your stained cell still alive? or you fixed?
if cell is still viable you can stain again with different antibody, no problem.
if you fixed the cell, prob not.

QUOTE (yftan2 @ Mar 5 2008, 05:32 PM)

I stained cells with different monoclonal antibodies. Unfortunately after staining the cells, our flow cytometer had broken down. I had the cells pulled and cryopreserved. I wonder would I be able to stain the cells with different monoclonal antibodies again after thawing them later?
Thx

thx a lot.